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Toxins 2019, 11(1), 35; https://doi.org/10.3390/toxins11010035

Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays

1
Postgraduate Programme in Pharmaceutical Sciences; Federal University of Santa Maria, Santa Maria 97105-900, Brazil
2
Industrial Pharmacy Department, Federal University of Santa Maria, Santa Maria 97105-900, Brazil
*
Author to whom correspondence should be addressed.
Received: 22 November 2018 / Revised: 28 December 2018 / Accepted: 10 January 2019 / Published: 12 January 2019
(This article belongs to the Special Issue Characterization and Quantitative Analysis of Botulinum Neurotoxin)
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Abstract

Botulinum neurotoxin type-A (BoNTA) is one of the seven different serotypes (A to G) produced by Clostridium botulinum. A stability-indicating size-exclusion chromatography (SEC) method was developed and validated, and the specificity was confirmed by forced degradation study, interference of the excipients, and peaks purity. The method was applied to assess the content and high-molecular-weight (HMW) forms of BoNTA in biopharmaceutical products, and the results were compared with those of the LD50 mouse bioassay, the T−47D cell culture assay, and the reversed-phase chromatography (RPC) method, giving mean values of 0.71% higher, 0.36% lower, and 0.87% higher, respectively. Aggregated forms showed significant effects on cytotoxicity, as well as a decrease in the bioactivity (p < 0.05). The employment of the proposed method in conjunction with the optimized analytical technologies for the analysis of the intact and altered forms of the biotechnology-derived medicines, in the correlation studies, enabled the demonstration of the capability of each one of the methods and allowed for great improvements, thereby assuring their safe and effective use. View Full-Text
Keywords: botulinum neurotoxin type A; size-exclusion chromatography; reversed-phase chromatography; T−47D cell culture; LD50 mouse bioassay botulinum neurotoxin type A; size-exclusion chromatography; reversed-phase chromatography; T−47D cell culture; LD50 mouse bioassay
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
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Xavier, B.; Perobelli, R.F.; Walter, M.E.; da Silva, F.S.; Dalmora, S.L. Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays. Toxins 2019, 11, 35.

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